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海水增养殖区监测技术规程

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附 录 C(规范性附录) 水利论文.e'K4^l,~C

@1x^"]%y0g y6i,H[0水中总磷测定—过硫酸钾氧化法
)Q be%m*C {qs|L0C.1 方法概述
~o8Ej,va^ZG0  将酸化的样品放于密封瓶中,用过硫酸盐热处理0.5h。实验表明如果样品是预先酸化处理的,氧化剂的用量可以从1%减少到0.3%。只简单地以相反顺序加磷酸盐试剂,所形成的游离氯被抗坏血酸还原。水利论文$I(pJ/l#p(L
C.2 采样和贮存水利论文D%?;^0G"t i(_1J0KD
  采样后尽快进行分析,最好在0.5h之内进行,不要超过2h。若不能立即进行分析,应在每100mL样品中加入1mL(4.5mol/L)硫酸酸化,放入聚乙烯瓶中在暗处冷贮存不超过24h。水利论文5@'D)jt;ZOy
C.3 器具水利论文zV$F*u(QO7j
  氧化反应在50mL硼硅玻璃瓶中进行,这种瓶具有金属夹固紧的玻璃塞。或用带螺旋盖的50mL瓶或锥形瓶。瓶子在使用之前用蒸馏水洗涤若干次。水利论文:UOvD[8p%C
  高压灭菌锅。水利论文7T*IX7OrR:U
C.4 试剂水利论文.G3O,~b,y+EvJ} c
  除GB17378.4中磷酸盐测定所列的试剂之外,还需要一种氧化溶液。水利论文O-D;R_2gE2g
  ——
V(BM,A3xv0过硫酸钾溶液Ⅰ:取15mL(4.5mol/L)硫酸,将5g过硫酸钾(分析纯)溶于此液中。将此饱和溶液在室温下贮存于聚乙烯瓶中,避免阳光直接照射。该溶液可稳定7d。
lT8Z#nMfO:r P8MTU0  —— 过硫酸钾溶液Ⅱ:将5g过硫酸钾溶解于100mL蒸馏水中。贮存方法同溶液Ⅰ。此溶液可稳定7d。水利论文uD`E:?!Z/Puw8}
C.5绘制工作曲线
^QUR6g|!`;q3J0  按GB17378.4中磷酸盐测定方法中相关要求配制磷酸盐标准溶液系列,注入氧化瓶中,加入2mL过硫酸钾溶液Ⅰ。盖紧瓶子,将其置于高压灭菌锅内,压热(120℃)处理30min。冷至室温后检查塞子是否密封(若不密封弃掉)。然后打开瓶盖,将溶液转入磷酸盐反应瓶中。其它分析步骤按制作磷酸盐工作曲线步骤进行。
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5P:uAml1D9`.f|0 水利论文| Bfz&o6\ v
附 录 D(规范性附录) 水利论文@,m4`*fs-[M0k

VOX F2p.G+ttK:p0沉积物总氮测定—次溴酸盐氧化法水利论文^d{ o[$I%T4[
D.1方法原理
]$[/Z"^3F ~0K;j"MS b0  样品在催化剂的作用下用浓硫酸消煮分解,使其中所含的氮转化成氨并与硫酸结合形成硫酸铵,在碱性介质中用次溴酸盐将其氧化为亚硝酸盐,然后以重氮—偶氮分光光度法测定。水利论文&^#I5mD$F
D.2 分析方法
*y!sU!L4M&l0D.2.1 仪器
fM;R)f.T |0  —— 50mL凯氏烧瓶若干;水利论文,F QPL5Y/W7IE{m
  —— 具塞50mL锥形瓶;
l)TT5N7y9@O7@0  —— 分光光度计;水利论文4V1U7ty.u P
  —— 小漏斗;
BW+k8o%s BoC2}0  —— 1000mL容量瓶。
q0U;I:KH(N0D.2.2试剂及配制水利论文qb0@ p2g0?/d0WU$y
  —— 无氨蒸馏水;水利论文R_ rrB%Zuv
  —— 浓硫酸;
U/Z u5u4fej'e0  —— 混合催化剂:硫酸钾与硫酸铜按10:1比例混合, 研细过80目筛,贮存于瓶中,消煮时每1mL浓硫酸加混合催化剂0.37g。
`/`"m7o.BL#Dv0  —— 铵标准贮备液 100mg/L-N
%E!X I*oJ(S-r0  称取0.4716g硫酸铵(预先在110℃下干燥1h)溶于少量水中,移入1000mL容量瓶中,加水至标线,混匀。再加1mL氯仿。贮存于棕色瓶中,冰箱中冷藏保存。此溶液1.00mL含氨-氮100.0μg,有效期为半年。水利论文#k6Eebs
  —— 铵标准使用液 10.0mg/L-N
1^ fJ"t `D0P/r%]q0移取10.0mL铵标准溶液于100mL容量瓶中,加水至标线,混匀。此溶液1.00mL含氨-氮10.0μg,每次使用前配制。
p w D+umo9L0  —— 氢氧化钠溶液 400mg/L水利论文0T4](}'A9~x
  称取200g氢氧化钠(优级纯)于1000mL水中,加热蒸发至500mL,盛于聚乙烯瓶中。水利论文.nq*JyI
  —— 盐酸溶液 1+1
R2~3tO I/JGAsP$J$D`0  500mL盐酸(密度=1.19g/mL)与同体积水混合而成。水利论文y5nV yb
  —— 溴酸钾-溴化钾贮备溶液水利论文E xbp E&J
  称取2.8g溴酸钾和20g溴化钾溶于1000mL水中,贮存于1000mL棕色瓶中。
s'_8R+g/Rmqh D.F0  —— 次溴酸钠溶液水利论文aeX*YZt]$}N
  量取1.0mL溴酸钾-溴化钾贮备溶液于250mL聚乙烯瓶中,加49mL蒸馏水和3.0mL盐酸溶液,盖紧摇匀,置于暗处5min后加入50mL氢氧化钠溶液,混匀。每次使用前配置。
d'X p"M-d1e:rM4r:h0  ——磺胺溶液 2g/L水利论文.J#ty ^1O k$Rb#@
  称取2.0g磺胺(NH2SO2C6H4NH2)溶于1000mL(1+1)盐酸溶液中,贮存于1000mL棕色瓶中。有效期为两个月。水利论文q-w"W#c@jl9m/W
  ——盐酸萘乙二胺溶液 1.0g/L水利论文:~.ga-O/O F
  称取0.50g盐酸萘乙二胺(C10H7NHCH2CH2NH2.2HCL)溶于500mL纯水中,贮存于棕色瓶中。冰箱中冷藏保存。有效期为一个月。
-@5K jU V0D.3 测定步骤
6v;^ N u yIA0D.3.1 绘制工作曲线
A6~ t!f0a+J5f%Q0  a)取6个100mL容量瓶分别加入0.00、0.10、0.20、0.40、0.60、0.80、1.00mL铵标准溶液,加水至标线,混匀。此时浓度分别为0.000、0.010、0.020、0.040、0.060、0.080、0.1000mg/L-N。水利论文8]6G(rEP"F'T ~
  b)各取25.0mL上述溶液分别置于50mL具塞锥形瓶中。
t C$Qx(Bn F~-QmY0  c)分别加入2.5mL次溴酸钠溶液,混匀,放置30min。再各加入2.5mL磺胺溶液,混匀,放置5min。最后各加入0.5mL盐酸萘乙二胺溶液混匀,放置15min。
:G*l7n,f2q/`I:P7G/\ p0  d)在543nm波长下,使用5cm比色皿,以无氨蒸馏水做参比测标准系列液的吸光度值Ai,其中0.00 mg/L-N浓度标准液所测吸光度值为Ao。水利论文"s&D:K X2wu6?
  e)以吸光度Ai-Ao为纵坐标,相应的浓度为横坐标,绘制工作曲线。
&aQU6E;O*i}H0D.3.2 样品测定
Wbmu` q?3_$G0  —— 称取1g待分析样品置于50mL凯氏烧瓶中,加1.85g混合催化剂和5mL浓硫酸, 水利论文(EtA+K5O+B0l)n
加盖小漏斗。开始用文火加热,待气泡发生后用大火,并不断摇动(硫酸在瓶内回流程度以高达瓶颈1/3处为好, 水利论文7T^s-~[ j[ f

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否则表示加热温度过高或过低),待溶液呈淡绿色后继续煮沸0.5h即可。稍冷后用蒸馏水将小漏斗内外洗净, 用约10mL蒸馏水溶解残渣(必要时可温热)。
IO#T5sx g!q-s/V{0  —— 将溶液定量转移到50mL锥形瓶中,依照D.3.1中a)~e)步骤测Aw。
/Ua:{[CMGY0  —— 不加样品而用25mL蒸馏水重复上两步骤测得空白吸光度值Ab。
Y,C#\8os3fW0  以Aw-Ab在工作曲线上查的样品中N的浓度。水利论文;m!p6L7OU{CZ B
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附 录 E(规范性附录)

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6Zi eb-@ k%q0沉积物总磷测定—Livingstone—Boykin法
n9T CFw h;a\!]0E.1 样品预处理
6K2MY%@%a3Hd0  —— 取适量沉积物样品于马弗炉中在450℃下焙烧6h;
HV-if-]$j R0  —— 称取焙烧沉积物样品0.1~0.5g于10mL聚四氟乙烯离心管中,加入H2SO4/HCLO4(V/V 为1/4)混合液4mL,
)AA&_,@U,x0在往复振荡器上振荡2h;水利论文#rw.N^p:ji+WY
  —— 上述萃取样品置于离心机中离心20min , 转速为3000rpm;
(V#h Z$E[ Z e e^x.R0  —— 取离心后的上清液2mL移入50mL具塞比色管中, 用重蒸馏水稀释至50mL。水利论文)i%y9d8gMK7b`
E.2 样品测定水利论文.W d w.S,H*I
  按GB17387.4海水中磷酸盐的测定方法,测量稀释液中的磷含量。
M9\wo(L,?a0 水利论文G^;p"R+C3ob
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附 录 F (规范性附录)

B2G3@{V S0x"?/]7h*}0 水利论文~(fPHz!A,H+{P"I+y

沉积物粪大肠杆菌—发酵法水利论文TO @7NZ JL
F.1 样品预处理
.H_7NC8x/D.D,p0  用无菌培养皿无菌称取1.0g(若沉积物是沙质,则样品量加倍,视具体情况而定)沉积物样品,然后无菌加入9.0mL(或加倍增加)无菌海水制备成10-1稀释样;再用无菌移液管将10-1稀释样连续稀释,制备成10-2、10-3、10-4稀释样。
#bB Ar C6n0F.2 分离方法水利论文_9V*V{-m6b
  按17387.9中规定的粪大肠杆菌检测方法进行检测。
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4r`+MxIu M0附 录 G (规范性附录) 水利论文5E]XBf&PP

水利论文,p@'y3DBe#x0Pj p

沉积物异养细菌总数—平板计数法
eMM I8K0G.1 样品预处理
u2}2?N6H]h8]W0  用无菌培养皿无菌称取1.0g(若沉积物是沙质,则样品量加倍,视具体情况而定)沉积物样品,然后无菌加入9.0mL(或加倍增加)无菌海水制备成10-1稀释样;再用无菌移液管将10-1稀释样连续稀释,制备成10-2、10-3、10-4稀释样。
m;@Gk(Eu0G.2 分离方法水利论文8o9Q BvreB
  按GB17387.7中规定的异养细菌总数检测方法进行检测。
"I!s @%j,p]!Y0 水利论文.a&aJ$a)p#{
 
F3RM%`q5t1{ _f;y%Y+o0附 录 H (规范性附录)

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m J0I#l1D+u^ v6o0海水增养殖区水质监测结果报表
7XG%m/]|0  海水增养殖区水质监测结果应按照表H1的内容计算、填报。水利论文,p_0Eg8b.{:j
 
o!wU?ef/X0附 录 I (规范性附录) 水利论文pY A#h4g

,l3V%yF!IXX0海水增养殖区沉积物监测结果报表
:x3|A4}a"]0  海水增养殖区沉积物监测结果应按照表I1的内容计算、填报。水利论文!V4kv^;AW egt J2a

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q$B5Dt3gz0附 录 J (规范性附录)

J*]0P6h5^:P+T+V%L0 水利论文lc/f?|/Z9f1j3Y

浮游植物细胞数量计数记录表水利论文5yJ[%{(N
  浮游植物细胞数量计数记录应按照表J1的内容计算、填报。

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CGgj3U/K,j,_0附 录 K (规范性附录) 水利论文:F2r nhX o w `

MJE&W[@zL0水质、沉积物质量评价报表
y&v A@ m)g/Pm"D0K.1 水质污染指数水利论文J8X};@.u.S
  水质各项污染因子的污染指数应按照表K1的内容计算、填报。水利论文"V,D4W3X0r:? l8[

/Q3J4u*aFr-@0水利论文-jr#i r nZ
附 录 L (资料性附录)

zI0q A hy+n3m"^0 水利论文k_fm*Tk*_.n

异养细菌总数评价等级
t,`-uCD)_cm0  异养细菌总数评价等级参见表L1。水利论文%G2`5H OCHD}m

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4vx{]j?)z!tY$q0附 录 M (规范性附录) 水利论文 r;m&\.CI0`

/Js Y ok{0海水增养殖区监测报告内容与格式
a}5L-G uyG0M.1 文本格式水利论文;pu9V'`G
M.1.1 文本规格水利论文0^nl't`K*I
  海水增养殖区监测报告文本外形尺寸为A4(210mm×297mm)。
8H n2Jf?i+~0M.1.2 封面格式
/[;Y#E+\`D W0  海水增养殖区监测报告封面格式如下。水利论文 {a.y9n?/[ K
  第一行书写:×××海区或省市×××海域(一号宋体,加黑,居中);水利论文0f5mI8CU?!LI Z+k$g1r
  第二行书写:海水增养殖区监测报告(一号宋体,加黑,居中);水利论文7k$d7o3iv5R B%L Y&x
  落款书写:编制单位全称(如有多个单位可逐一列入,三号宋体,加黑,居中);
0z rr,Yz"M+uve6`m0  第四行书写:××××年××月(小三号宋体,加黑,居中);水利论文v#l c.}[ iC FHXi
  第五行书写:中国,空一格,××(地名,小三号宋体,加黑,居中);水利论文1C?:V;h9x\J
  以上各行间距应适宜,保持封面美观。水利论文0SvE"Z;md$d%R {n'v
M.1.3 封里一内容水利论文 h Xy!\t5T|(q X
  封里一中应分行写明:监测项目实施单位全称(加盖公章);项目负责人、技术总负责人、分项目负责人(包括生物因子、水化学因子等分项)和主要参加人员姓名;报告书编制单位全称(加盖公章);编制人、审核人姓名;编制单位地址;通信地址;邮政编码;联系人姓名;联系电话;E-mail地址等内容。水利论文 bLf*V`'b)m
M.2 海水增养殖区监测报告章节内容
0z}1z-p5?[:pqg%c0  海水增养殖区监测报告应包括以下全部或部分内容。依据监测目的、内容和具体要求,可对下列章节及内容适当增减。水利论文+N.UUe'}
  前言水利论文$Zl$mk!lmY!t_c
  主要包括海水增养殖区监测工作任务来源、监测任务实施单位、养殖区概况(养殖种类、方式、数量、规模等)、监测时间与航次、监测船只与合作单位等的简要说明。水利论文}c2[0pC,me(?4[
  1 监测方案水利论文bN$^]3np
    1.1 监测海区的区域与范围
9V {*]9M^Qz3G0    1.2 监测站位布设
,~ \R@a8m2Y0    1.2.1 监测站位图水利论文,OICA*_:]:mW
    1.2.2 监测站位类型与说明
1r_IXF:F ^2fC0    1.2.3 监测时间与频率
M\._ oGJ3SGC0  2 监测内容与检测分析水利论文.f\{$Hu B$Z'wQ
    2.1 水质监测与分析结果
w V G v(h9q0    2.1.1 水文要素
].X{4{k1m&V"h2xW0       主要包括水温、透明度、盐度等项目的监测与分析结果。水利论文.pPY*t6v*B:hr6Tsgeu
    2.1.2 化学要素
h2VYCW0       主要包括汞、镉、铅、锌、砷、油类、磷酸盐、硅酸盐、氨氮、硝酸盐、亚硝酸盐、溶解氧、化学需氧量、pH等项目的监测与分析结果。水利论文*V2t"NT+t7?8n
    2.1.3 生物要素
N*c$Hgr[9v%p0      主要包括粪大肠杆菌、弧菌数量、异养细菌总数、叶绿素a含量、浮游植物的种类与数量等生物学指标的监测与分析结果。
j2Q r{;f0    2.2 沉积物监测与分析结果(加*号为选测项目)水利论文m uKL-e1{m
    2.2.1 化学要素水利论文k7| L$V B Z
      主要包括汞、镉、铅、锌、砷、油类、DDT、PCBs、有机质、硫化物等项目的监测与分析结果。水利论文&s?;C.xsD'KEG9]+V
    2.2.2 生物要素
Pt*Zjz0       主要包括粪大肠杆菌、弧菌数、异养细菌总数等生物学指标的监测与分析结果。
Q0{*C+R ~&GI u)U0  3 分析与评价
iD+r.Y%yir0    3.1 污染指数指标
{-U!CP6EE"G4RSw0      主要包括海水和沉积物的化学要素的单因子污染指数分析评价。
h|RY|GEY0    3.2 浮游植物生物多样性指标水利论文0Xkvi&T
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   3.3 营养水平指标水利论文"eY$c&{\"O J3Jq
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  4 海水增养区监测结果的综合分析与评价
L"vv4d hR5n0   4.1 监测海区的水环境质量现状的分析与评价
5yyQQTSAe0   4.2 监测海区的沉积物环境的分析与评价水利论文%SixO7]0r
   4.3 监测海区环境质量发展趋势的分析与预测
k(v(f&[$pI0   4.4 对监测海区环境质量管理的措施与建议
8e~*f;W.u6?U)W:{d d0  5 监测数据报表
~5lp"_Dq LA0   水质、沉积物、浮游植物等监测结果汇表。水利论文` J\.F(~ V}zK
  6 附图、附表、附件
{Xa^-@9}0   含参考文献、等值线图等。
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  本规程的附录A、附录B、附录C、附录D、附录E、附录F、附录G、附录H、附录I、附录J、附录K和附录M为规范性附录,附录L为资料性附录。
] uU c_"Np;xh0  本规程由国家海洋局海洋环境保护司第一次提出。
BMwl7S.c0  本规程起草单位:国家海洋环境监测中心。
+O/v%x g q0?*V0  本规程主要起草人:王立俊、王健国、梁玉波、韩庚辰。

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