《芒果果实中有机酸分析方法 高效液相色谱法》编制说明（征求意见稿）

《芒果果实中有机酸分析方法高效液相色谱法》编制说明（征求意见稿）根据《广西农业农村产业振兴促进会关于<芒果果实中有机酸测定高效液相色谱法>等6项团体标准立项的通知》文件要求，本标准由广西壮族自治区农GB5009.157-2016食品安全国家标准食品NB/T12004-2016费托合成水中有机酸、醇、醛、酮的NY/T2277-2012水果蔬菜中有机酸和阴离子的测定离子色500-2014烟草及烟草制品挥发性有机酸的测定气相色谱-质谱8657-2014苯乙烯-丁二烯生橡胶T/NAIA0200-2023葡萄酒中多种有机酸的测定T/NAIA0207-2023酿酒葡萄及葡萄酒中有机酸的测定液T/NAIA098-2021酿酒葡萄及葡萄酒中有机酸含量的测定的侧定方法。适用于果汁及果汁饮料、碳酸饮料、固体饮料、胶基糖果、饼干、经查阅,国内外未制定有与利用高效液相色谱法分析芒果果实中有机酸的相本标准根据GB/T1.1—2020《标准化工作导则第1部分：标准化文件的结％）准储备液；a-酮戊二酸、富马酸、没食子酸标准品40.00mg用纯净水定容至10酸、柠檬酸、琥珀酸配置成浓度为800mg/L；草酸配置成浓度为400），13.19mg/kg、64.01～1541.38mg/kg、2.59～133.97mg/kg。柠檬酸、乳酸、图1芒果果实中11种有机酸物质含量4高效液相色谱法测定芒果中草酸、酒石酸、奎宁酸、苹果酸、a-酮戊二在190～220nm之间的波长。这是因为有机酸（如乙酸、乳酸、柠檬酸等）通常在紫外光谱的远紫外区具有吸收峰。这一区域的吸收主要与有机酸的羧基（-COOH）基团相关。190nm适合检扫描波长范围190～300nm，对草酸、酒石酸、奎宁酸、乳酸、乙酸、柠檬酸、富马酸、琥珀酸、没食子酸分别进行光谱扫描。由图2明Shim-packGISC18峰型偏圆，PerfectT3峰型较好，能将有机酸分离开来进一步对流动相流速进行优化，选用0.01mol/LpH2.8磷酸二氢钾中含3%行等度洗脱。结果表明流动相流速为0.5ml/min时，有机酸出峰效果最好（图在进行方法准确度与精密度之前，首先对全国40份芒果样品进行检测，发琥珀酸、没食子酸含量最多582.40mg/kg14.83mg/kg、1731.58mg/250.14～571.40mg/kg，314.63～2161.58mg/kg，248.41～2211.27～2524.91mg/kg，0～120.95mg/kg，237.71～3187.08mg/kg，78.90~0～133.97mg/kg。设置25mg/kg、250mg/k计算回收率，确定方法准确性及精密度，结果见表2。组分试样组分浓度（mg/kg）理论加标浓度（mg/kg）加标试样组分浓度（mg/kg）回收率(%)平均回收率(%)RSD(%)草酸2534.7298.2095.912.3033.5793.6034.1595.9225035.99103.28103.320.1836.05103.5235.96103.16500528.10103.5996.076.53469.5791.88473.8292.73酒石酸2542.29106.00105.810.6242.37106.3242.07250287.86108.83101.886.03262.7298.77260.9298.05500518.09100.46102.452.95545.00105.84521.05101.05奎宁酸2538.03102.44104.604.2039.78109.4437.90101.92250268.77102.54102.213.18259.6098.87275.44105.21500540.00105.51107.492.17561.52109.82548.15107.15苹果酸50.522573.6692.5694.6374.7296.8074.1594.52250279.6298.8498.570.27296.2798.30296.9498.57500562.07102.31102.720.42566.22102.72a-酮戊二酸6.052530.2796.8893.922.9628.7990.9639.5393.92250257.86100.72100.312.45250.2597.68262.40102.54500546.55108.10107.540.55541.00106.99543.80107.55乳酸35.312562.25107.76105.572.0961.21103.6061.65105.36250305.86108.22106.702.85306.50108.48293.83103.41500570.31107.00104.262.58544.66101.87554.86103.9125.202550.0299.2897.3649.0695.4449.5497.36250270.7298.2198.690.448373.1599.18271.9298.69500536.25102.21103.70104.15548.80104.72柠檬酸195.8625221.84103.92104.363.16221.22101.44222.79107.72250461.31106.18105.440.74457.63104.71459.46105.44500706.81102.19102.130.07102.05706.56富马酸2.612528.96105.400.7229.32106.84250263.52104.36103.520.84259.30102.68261.40103.52500507.40100.96102.01517.95103.07512.65102.01琥珀酸49.912574.1596.9698.2074.7799.4474.4698.20250293.4597.4296.640.89292.0096.84289.0895.67500501.0690.2392.882.33518.7193.76523.1094.64没食子酸3.332528.2099.4899.770.4428.40100.2828.2299.56250260.33102.80101.93258.78102.18255.35100.81500527.05104.74103.890.84518.60103.05522.80103.89组分浓度（mg/L）峰面积RSD草酸80.005739975790345765150.44酒石酸400.004207684130684169180.92奎宁酸800.00494747508533501640苹果酸800.005423725385485404600.35a-酮戊二酸3601823562533582170.55乳酸800.003227533188973208250.60800.003799283735803767540.84柠檬酸800.00694231675359684795富马酸135205013230191337535琥珀酸800.00398647386644392645没食子酸188081718248031852810稀释梯度下有机酸的色谱图，确定方法的校准曲线及相关系数，结果见表3。序号组分线性范围（mg/L）线性方程相关系数R21草酸3.125~400y=7710x-9540.60.99922酒石酸6.25~800y=1801.4x-1631.10.99933奎宁酸6.25~800y=191.08x+4791.50.99994苹果酸6.25~800y=739.24x+290.790.99975a-酮戊二酸y=1796.25x-1585.60.99896乳酸6.25~800y=382.81x-509.860.999376.25~800y=509.53x-657.240.99958柠檬酸6.25~800y=811.76x+285.220.99909富马酸y=10416x-1573.50.9997琥珀酸6.25~800y=516.21x-1050.50.9997没食子酸y=12763x-4730.90.9997果果实中有机酸的平均回收率在92.88107.54％之间，相对标准偏差在0.076.53％之间，10种有机酸在液相色谱仪上响应值的RSD为0.44%~酸检出限为3.125mg/kg，定量限为6.25酸检出限为3.125mg/kg，定量限为6.25的检测方法处理样品，计算回收率。试验结果表明，25mg/kg、250mg组分(%)(%)RSD(%)草酸38.632561.7592.4894.522.0662.7296.3662.3194.72250270.5392.7695.953.00280.4396.72284.5398.36500533.0398.8899.96537.7099.81544.54101.18酒石酸33.602559.49103.56105.2960.08105.9260.20106.40250290.05102.58105.222.64296.00104.96303.87500525.3598.3594.233.81496.5592.59492.4091.76奎宁酸42.582565.7292.5695.432.6466.9197.3266.6896.40250313.98108.56102.155.59293.30100.29286.5897.60500554.44102.37105.393.52590.25109.53563.90104.26苹果酸211.5325234.4291.5694.233.72236.0898.20234.7992.92250460.0799.42104.414.42471.75105.29482.83108.52500739.98105.69107.90761.28109.95751.86108.07a-酮戊二酸8.562531.4391.4892.934.0031.1090.1632.8597.16250275.62106.822.28266.78103.29264.32102.30500522.60102.81101.212.28519.91102.27501.3598.56乳酸20.682545.3298.5695.314.5044.9196.9243.2990.44250277.83102.86104.592.20288.68107.20279.99103.72500530.15101.89102.062.47518.8199.63543.97104.6636.492562.87105.5297.077.9559.0990.4060.3195.28250282.4498.3899.293.9995.86295.55103.62500544.47101.60104.472.39106.13564.91105.68柠檬酸30.4325106.7698.957.2853.5792.5654.8197.5225098.2899.23281.83100.56277.5998.86500101.74104.392.99548.38103.59569.57107.83富马酸5.982530.8199.3298.634.6231.68102.8029.4293.76250266.67104.28104.27263.46102.99269.81105.53500547.93108.39104.573.27523.46103.50515.04101.81琥珀酸32.002557.86103.4495.6554.7190.8455.1792.68250277.9998.40102.093.92297.86106.34285.85101.54500571.97107.99107.940.61568.27107.25574.86108.54没食子酸2538.1594.6095.844.7039.71100.8437.5292.08250252.8195.3295.454.91241.5990.83265.03100.21500535.76104.25102.432.04528.97102.89515.0组分试样组分浓度（mg/kg）理论加标浓度（mg/kg）加标试样组分浓度（mg/kg）回收率(%)平均回收率(%)RSD(%)草酸34.962561.67106.84104.344.0359.8399.4861.64106.72250291.26102.5299.183.92272.2494.91285.25500549.58102.92100.702.08537.02100.41528.7598.76酒石酸39.732564.4298.76100.2565.27102.1664.6999.84250295.37102.26100.49289.79100.02287.6999.18500540.13100.08100.430.66544.72101.00545.75101.20奎宁酸25.552550.2198.64100.645.4152.25106.8049.6796.48250269.0597.4096.27260.7294.07268.9897.37500528.75100.64100.290.39527.41100.37524.8499.86苹果酸32.602559.19106.36105.9659.44107.3658.64104.16250283.21100.24100.07285.56101.18297.5798.79500532.0999.9099.820.40533.44100.17529.4999.38a-酮戊二酸6.742530.6595.6499.493.4632.31102.2831.88100.56250263.72102.79102.700.08263.33102.64263.41102.67500509.81100.61100.500.10509.13100.48508.74100.40乳酸2534.2598.0894.983.4234.8095.2833.8891.60250261.89100.36100.040.66259.2999.32261.50100.21500513.80100.5699.710.81505.4998.90508.9899.602538.7595.5693.352.5237.5890.8838.2693.60250277.24104.9596.427.81241.5890.69248.9493.63500532.90103.61102.480.71528.70102.77525.60102.15柠檬酸25.432550.77101.3697.923.3449.8297.5694.84250275.80100.1598.272.48264.2195.51273.3099.15500527.00100.31103.903.08550.30104.97557.60106.43富马酸9.282534.83102.2098.993.3634.0899.2033.1795.5625099.94101.532.09103.94261.03100.70500509.0399.95100.000.04509.33100.01509.47100.04琥珀酸2545.97106.20101.285.4743.2495.2845.01102.36250273.88101.78101.39275.87102.58268.9599.81500514.2898.9799.670.61519.53100.02519.50100.02没食子酸5.802528.4890.7299.448.3130.91100.4432.59107.16250252.7598.7899.620.73256.07255.6999.96500100.06100.000.06505.7799.99505.5299.94组分试样组分浓度（mg/kg）理论加标浓度（mg/kg）加标试样组分浓度（mg/kg）回收率(%)平均回收率(%)RSD(%)草酸42.722568.50105.252.3468.89104.6869.71107.96250294.95100.89103.562.73300.86103.26309.02106.52500553.68102.19102.720.47558.9103.13556.96102.85酒石酸80.4425103.8293.5297.357.90106.99106.20103.5292.32250340.72105.60343.79105.34348.83107.36500592.35102.38101.730.97591.51102.21583.38100.59奎宁酸33.942562.9796.414.4564.78101.3662.8893.76250298.86103.69102.830.89294.08101.86296.80102.94500550.50102.21103.76564.25104.96560.05苹果酸23.212549.38104.68106.79107.6450.22108.04250262.3095.6495.65267.4097.68257.3093.64500523.86100.130.10524.19100.20523.21100.00a-酮戊二酸4.582530.3696.596.2328.4295.3627.4091.28250249.0297.7996.52242.7595.27245.8096.49500500.0299.0999.580.43503.9499.87503.5299.79乳酸2535.78101.88101.002.1935.97102.6434.9398.48250259.5499.6999.890.56261.61100.52258.9399.45500516.25101.19101.290.15516.36101.21517.63101.462543.8795.80103.316.4046.99108.2846.38105.84250278.70103.51103.560.18278.42103.40279.32103.76500515.5599.1399.340.28518.2099.66516.1399.24柠檬酸2570.55105.72106.0470.28104.6471.06107.76250306.20104.83103.780.97303.30103.67301.20102.83500545.50100.2899.81546.80100.54537.1098.60富马酸8.702534.91104.84101.053.5633.86100.6497.68250257.9799.7198.40255.8798.87250.2396.61500508.2099.9098.910.99498.3397.93503.1998.90琥珀酸9.482532.9593.8896.765.1932.9493.84102.56250263.90101.77103.72269.78272.66105.27500519.40101.98101.600.41515.22101.15517.85101.67没食子酸6.432530.2795.3695.370.2330.2295.1630.3395.60250261.62102.08102.050.56262.96102.61206.09101.46500506.66100.05100.220.17508.42100.40507.47100.21组分试样组分浓度（mg/kg）理论加标浓度（mg/kg）加标试样组分浓度（mg/kg）回收率(%)平均回收率(%)RSD(%)草酸47.772573.55102.8797.4874.77108.00250305.550.34306.52103.50304.77102.80500578.85106.22106.000.26576.16105.68578.22106.09酒石酸54.432580.75105.28104.1380.62104.7680.02102.36250307.51101.23100.10306.25100.73300.2698.33500551.0299.3298.940.44549.63199.04546.7998.47奎宁酸49.612593.233.7473.8697.0072.7592.56250301.21100.64100.950.31301.96100.94302.75101.26500549.81100.04100.630.55555.32553.17100.71苹果酸77.7525104.77108.08106.912.27104.88108.52103.78250331.89101.66103.20337.80104.02337.52103.91500577.5299.9510